An Efficient Method for the Expression and Purification of Aβ(M1–42)
| Autor: | Sheng Zhang, Stan Yoo, Adam G. Kreutzer, James S. Nowick |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
chemistry.chemical_classification Amyloid beta-Peptides Gene Expression Peptide 010402 general chemistry Cleavage (embryo) 01 natural sciences Biochemistry Peptide Fragments Recombinant Proteins Article 0104 chemical sciences law.invention 03 medical and health sciences 030104 developmental biology Enzyme chemistry law Escherichia coli Recombinant DNA Humans |
| Zdroj: | Biochemistry. 57:3861-3866 |
| ISSN: | 1520-4995 0006-2960 |
| Popis: | Advances in amyloid research rely on improved access to the β-amyloid peptide, Aβ. N-terminal methionine extended Aβ, Aβ(M1–42), is a readily expressed and widely used form of Aβ with comparable properties to the natural Aβ(1–42) peptide. Expression of Aβ(M1–42) is simple to execute and avoids an expensive and often difficult enzymatic cleavage step associated with expression and isolation of Aβ(1–42). This paper reports an efficient method for expression and purification of Aβ(M1–42) and (15)N-labeled Aβ(M1–42). This method affords the pure peptide at about 19 mg per liter of bacterial culture through simple and inexpensive steps in three days. This paper also reports a simple method for construction of recombinant plasmids, and the expression and purification of Aβ(M1–42) peptides containing familial mutations. We anticipate that these methods will enable experiments that would otherwise be hindered by insufficient access to Aβ. |
| Databáze: | OpenAIRE |
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